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Baetscher, Manfred

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Baetscher

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Manfred

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Baetscher, Manfred
Author's Publications: search.filters.isAuthorOfPublication.6f841a24-2973-4acc-be74-017ff9c611f6

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    Reprogramming within Hours Following Nuclear Transfer into Mouse but not Human Zygotes
    (Nature Publishing Group, 2011) Egli, Dieter; Chen, Alice E.; Saphier Belfer, Genevieve; Ichida, Justin; Fitzgerald, Claire; Go, Kathryn J.; Acevedo, Nicole; Patel, Jay; Baetscher, Manfred; Kearns, William G.; Goland, Robin; Leibel, Rudolph L.; Melton, Douglas; Eggan, Kevin
    Fertilized mouse zygotes can reprogram somatic cells to a pluripotent state. Human zygotes might therefore be useful for producing patient-derived pluripotent stem cells. However, logistical, legal and social considerations have limited the availability of human eggs for research. Here we show that a significant number of normal fertilized eggs (zygotes) can be obtained for reprogramming studies. Using these zygotes, we found that when the zygotic genome was replaced with that of a somatic cell, development progressed normally throughout the cleavage stages, but then arrested before the morula stage. This arrest was associated with a failure to activate transcription in the transferred somatic genome. In contrast to human zygotes, mouse zygotes reprogrammed the somatic cell genome to a pluripotent state within hours after transfer. Our results suggest that there may be a previously unappreciated barrier to successful human nuclear transfer, and that future studies could focus on the requirements for genome activation.
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    An Embryonic Stem Cell-Based System for Rapid Analysis of Transcriptional Enhancers
    (Wiley Periodicals, Inc., 2012) Tsanov, Kaloyan M; Nishi, Yuichi; Peterson, Kevin A; Liu, Jing; Baetscher, Manfred; McMahon, Andrew P.
    With the growing use of genome-wide screens for cis-regulatory elements, there is a pressing need for platforms that enable fast and cost-effective experimental validation of identified hits in relevant developmental and tissue contexts. Here, we describe a murine embryonic stem cell (ESC)-based system that facilitates rapid analysis of putative transcriptional enhancers. Candidate enhancers are targeted with high efficiency to a defined genomic locus via recombinase-mediated cassette exchange. Targeted ESCs are subsequently differentiated in vitro into desired cell types, where enhancer activity is monitored by reporter gene expression. As a proof of principle, we analyzed a previously characterized, Sonic hedgehog (Shh)-dependent, V3 interneuron progenitor (pV3)-specific enhancer for the Nkx2.2 gene, and observed highly specific enhancer activity. Given the broad potential of ESCs to generate a spectrum of cell types, this system can serve as an effective platform for the characterization of gene regulatory networks controlling cell fate specification and cell function.